o-Vanillin binds covalently to MAL/TIRAP Lys-210 but independently inhibits TLR2.

Toll-like receptor (TLR) innate immunity signalling protects against pathogens, but excessive or prolonged signalling contributes to a range of inflammatory conditions. Structural information on the TLR cytoplasmic TIR (Toll/interleukin-1 receptor) domains and the downstream adaptor proteins can help us develop inhibitors targeting this pathway. The small molecule o-vanillin has previously been reported as an inhibitor of TLR2 signalling. To study its mechanism of action, we tested its binding to the TIR domain of the TLR adaptor MAL/TIRAP (MALTIR). We show that o-vanillin binds to MALTIR and inhibits its higher-order assembly in vitro. Using NMR approaches, we show that o-vanillin forms a covalent bond with lysine 210 of MAL. We confirm in mouse and human cells that o-vanillin inhibits TLR2 but not TLR4 signalling, independently of MAL, suggesting it may covalently modify TLR2 signalling complexes directly. Reactive aldehyde-containing small molecules such as o-vanillin may target multiple proteins in the cell.

Geographical origin discriminatory analysis of onions: Chemometrics methods applied to ICP-OES and ICP-MS analysis.

Geographical origin is an important determinant of agricultural product quality and safety. Herein, inductively coupled plasma (ICP) analysis was applied to determine the inorganic elemental content of onions and identify their geographical origin (Korean or Chinese). Chemometric, including principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and orthogonal partial least square discriminant analysis (OPLS-DA) were applied to the ICP results. OPLS-DA distinguished each group, and 17 elements with variable importance in projection (VIP) values of ≥ 1 were selected. The receiver operating characteristic (ROC) curve had an area under the curve (AUC) of 1, indicating excellent discriminatory power. Differences in elemental content between groups were visually observed in a heatmap, and the country of origin was determined with 100% accuracy using canonical discriminant analysis (CDA). This method accurately distinguishes between Korean and Chinese onions and is expected to be beneficial for identifying agricultural products.

ONECUT2 Activates Diverse Resistance Drivers of Androgen Receptor-Independent Heterogeneity in Prostate Cancer.

Androgen receptor- (AR-) indifference is a mechanism of resistance to hormonal therapy in prostate cancer (PC). Here we demonstrate that the HOX/CUT transcription factor ONECUT2 (OC2) activates resistance through multiple drivers associated with adenocarcinoma, stem-like and neuroendocrine (NE) variants. Direct OC2 targets include the glucocorticoid receptor and the NE splicing factor SRRM4, among others. OC2 regulates gene expression by promoter binding, enhancement of chromatin accessibility, and formation of novel super-enhancers. OC2 also activates glucuronidation genes that irreversibly disable androgen, thereby evoking phenotypic heterogeneity indirectly by hormone depletion. Pharmacologic inhibition of OC2 suppresses lineage plasticity reprogramming induced by the AR signaling inhibitor enzalutamide. These results demonstrate that OC2 activation promotes a range of drug resistance mechanisms associated with treatment-emergent lineage variation in PC. Our findings support enhanced efforts to therapeutically target this protein as a means of suppressing treatment-resistant disease.

Ultra-High-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry for Simultaneous Pesticide Analysis and Method Validation in Sweet Pepper.

Pesticides effectively reduce the population of various pests that harm crops and increase productivity, but leave residues that adversely affect health and the environment. Here, a simultaneous multicomponent analysis method based on ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) pretreated by the QuEChERS method was developed to control the maximum residual levels. Among the 140 pesticides with high frequency of detection in agricultural products in Gyeongnam region in Korea for 5 years, 12 pesticides with high detection frequency in sweet pepper were selected. The analytical method is validated, linearities are r2 > 0.999, limit of detection (LOD) ranges from 1.4 to 3.2 µg/kg, and limit of quantification (LOQ) ranges from 4.1 to 9.7 µg/kg, and the recovery rate was 81.7-99.7%. In addition, it was confirmed that a meaningful value of these parameters can be achieved by determining the measurement uncertainty. The results proved that parameters such as recovery rate and relative standard deviation of the analysis method were within international standards. Using the developed method, better and safer sweet peppers will be provided to consumers, and effective pesticide residue management will be possible by expanding to other agricultural products.

Development of a method for analysis and risk assessment of residual pesticides in ginseng using liquid and gas chromatography-tandem mass spectrometry.

In this study, we developed a method for detecting 335 pesticides in ginseng using liquid chromatography quadrupole mass spectrometry (LC-MS/MS) and gas chromatography quadrupole mass spectrometry (GC-MS/MS). Additionally, the linearity, sensitivity, selectivity, accuracy, and precision of the method was validated. The limits of detection (LOD) and limits of quantification (LOQ) for the instrument used in these experiments was 0.1-5.8 µg/kg and 0.3-17.5 µg/kg, respectively. The average recovery was 71.6-113.4%. From 2016 to 2019, 467 ginseng samples were analyzed, of which 304 samples detected pesticide residues, but most of them were below the standard. It can be observed that the hazard quotient (HQ) of ginseng for detected pesticides was less than 1, thus implying that the risk was low. Hence, in this study, we developed a specific, reliable, and suitable method for a fast and simultaneous analysis of 335 pesticides in ginseng.

Geographical discrimination of dried chili peppers using femtosecond laser ablation-inductively coupled plasma-mass spectrometry (fsLA-ICP-MS).

This study presents a method for discriminating the geographical origin of dried chili peppers using femtosecond laser ablation-inductively coupled plasma-mass spectrometry (fsLA-ICP-MS) and multivariate analysis, such as orthogonal partial least squares discriminant analysis (OPLS-DA), heatmap analysis, and canonical discriminant analysis (CDA). Herein, 102 samples were analyzed for the content of 33 elements using optimized conditions of 200 Hz (repetition rate), 50 µm (spot size), and 90% (energy). Significant differences in count per second (cps) values of the elements were observed between domestic and imported peppers, with variations of up to 5.66 times (133Cs). The OPLS-DA model accuracy achieved an R2 of 0.811 and a Q2 of 0.733 for distinguishing dried chili peppers of different geographical origins. The variable importance in projection (VIP) and s-plot identified elements 10 and 3 as key to the OPLS-DA model, and in the heatmap, six elements were estimated to be significant in discriminating between domestic and imported samples. Furthermore, CDA showed a high accuracy of 99.02%. This method can ensure food safety for consumers, and accurately determine the geographic origin of agricultural products.

Discrimination between Korean and Chinese Kimchi using inductively coupled plasma-optical emission spectroscopy and mass spectrometry: A multivariate analysis of Kimchi.

Kimchi has been designated as one of the world's five healthiest foods, and it is a traditional Korean fermented food. The purpose of this study was to investigate whether the origin of Kimchi can be discriminated against by using inorganic elements to develop a more accurate method. The OPLS-DA showed that the R2 and Q2 values were 0.908 and 0.81, a high-quality model. We selected 24 elements (133Cs, 238U, 88Sr, K, 157Gd, Na, Mg, 139La, P, 141Pr, 72Ge, 146Nd, 147Sm, 153Eu, 55Mn, 165Ho, 163Dy, 166Er, Fe, 172Yb, 169Tm, 185Re, 175Lu, and 118Sn) with VIP 1 or higher in the OPLS-DA model. In ROC, the selected elements had an accuracy of 100%. The heatmap confirmed the contents of rare earth elements (REEs) in Korean and Chinese Kimchi, the accuracy of distinguishing classification in CDA is 100%. Such results will help to distinguish the origin of agricultural products through inorganic analysis.

Correlation between Collagen Type I/III Ratio and Scar Formation in Patients Undergoing Immediate Reconstruction with the Round Block Technique after Breast-Conserving Surgery.

The aim of this study was to investigate the relationship between the collagen type I/III ratio and scarring in patients who underwent immediate reconstruction with the round block technique (RBT) after breast conservation surgery. Seventy-eight patients were included, and demographic and clinical characteristics were recorded. The collagen type I/III ratio was measured using immunofluorescence staining and digital imaging, and scarring was assessed using the Vancouver Scar Scale (VSS). The mean VSS scores were 1.92 ± 2.01 and 1.79 ± 1.89, as assessed by two independent plastic surgeons, with good reliability of the scores. A significant positive correlation was found between VSS and the collagen type I/III ratio (r = 0.552, p < 0.01), and a significant negative correlation was found between VSS and the collagen type III content (r = -0.326, p < 0.05). Multiple linear regression analysis showed that the collagen type I/III ratio had a significant positive effect on VSS (ß = 0.415, p = 0.028), whereas the collagen type I and collagen type III content had no significant effect on VSS. These findings suggest that the collagen type I/III ratio is associated with scar development in patients undergoing RBT after breast conservation surgery. Further research is needed to develop a patient-specific scar prediction model based on genetic factors affecting the collagen type I/III ratio.

Comparison of CT- and MRI-Based Quantification of Tumor Heterogeneity and Vascularity for Correlations with Prognostic Biomarkers and Survival Outcomes: A Single-Center Prospective Cohort Study.

BACKGROUND: Tumor heterogeneity and vascularity can be noninvasively quantified using histogram and perfusion analyses on computed tomography (CT) and magnetic resonance imaging (MRI). We compared the association of histogram and perfusion features with histological prognostic factors and progression-free survival (PFS) in breast cancer patients on low-dose CT and MRI. METHODS: This prospective study enrolled 147 women diagnosed with invasive breast cancer who simultaneously underwent contrast-enhanced MRI and CT before treatment. We extracted histogram and perfusion parameters from each tumor on MRI and CT, assessed associations between imaging features and histological biomarkers, and estimated PFS using the Kaplan-Meier analysis. RESULTS: Out of 54 histogram and perfusion parameters, entropy on T2- and postcontrast T1-weighted MRI and postcontrast CT, and perfusion (blood flow) on CT were significantly associated with the status of subtypes, hormone receptors, and human epidermal growth factor receptor 2 (p < 0.05). Patients with high entropy on postcontrast CT showed worse PFS than patients with low entropy (p = 0.053) and high entropy on postcontrast CT negatively affected PFS in the Ki67-positive group (p = 0.046). CONCLUSIONS: Low-dose CT histogram and perfusion analysis were comparable to MRI, and the entropy of postcontrast CT could be a feasible parameter to predict PFS in breast cancer patients.

Flow cytometric reporter assays provide robust functional analysis of signaling complexes.

Conventional assays to probe signaling protein interactions and function involve measurement of luciferase reporter expression within the bulk cell population, with lack of control over target-protein expression level. To address this issue, we have developed a rapid and robust flow cytometric assay for analysis of signaling protein function. A fluorescent reporter and fluorescent tagging of the target protein enables simultaneous assessment of protein expression and signaling within individual cells. We have applied our technique to the analysis of variants of the lipopolysaccharide receptor Toll-like receptor 4 (TLR4) and its adapter protein MyD88, using a NF-кB-responsive promoter driving mScarlet-I expression. The assay enables exclusion of nontransfected cells and overexpressing cells that signal spontaneously. Additionally, our assay allows the identification of protein variants that fail to express. We found that the assays were highly sensitive, with cells expressing an appropriate level of GFP-MyD88 showing approximately 200-fold induction of mScarlet-I by lipopolysaccharide, and we can detect subtle protein concentration-dependent effects of mutations. Importantly, the assay is adaptable to various signaling pathways.

"Tear-Drop Appearance" Wrap: A Novel Implant Coverage Method for Creating Natural Contour in Prepectoral Prosthetic-Based Breast Reconstruction.

Various implant wrapping methods with acellular dermal matrix (ADM) have been introduced, but most focus on random trimming and suturing aimed to maximize implant coverage. Here we present our clinical experience using a "tear-drop appearance" wrapping method to achieve natural contours through upper pole volume replacement. We retrospectively reviewed the data of 56 consecutive cases of prepectoral prosthetic-based breast reconstruction (PPBR) using this wrapping method following nipple-sparing mastectomy between March 2020 and June 2021. The "tear-drop appearance" wrapping design creates an anatomical tear-drop-shaped pocket to encourage lower pole fullness and create a natural contour through upper pole volume replacement by ADM. Patients' baseline characteristics, operative data, and complications were analyzed. Aesthetic outcomes were measured using the BREAST-Q and Aesthetic Item Scale (AIS). A successful reconstruction was achieved without major complications and using a single ADM sheet. Four types and three sizes of ADMs were used. The mean resected breast tissue weight was 274.3 g, while the mean implant volume was 230.0 cc. The average BREAST-Q and AIS scores were 4.6 ± 0.8 and 4.5 ± 0.7, respectively. Owing to its simplicity, reproducibility, and effectivity, this method is an excellent implant coverage option that achieves a natural contour in PPBR.

Development of a Quantitative Method for Detection of Multiclass Veterinary Drugs in Feed Using Modified QuPPe Extraction and LC-MS/MS.

A method was developed for the rapid and quantitative analysis of 30 veterinary drugs belonging to 17 classes (amphenicols (1), anthelmintics (1), cephalosporins (4), coccidiostats (1), lincosamides (1), macrolide (1), nitroimidazole (1), penicillins (3), phenylhydrazines (1), polypeptides (1), pyrethrins (1), quinolones (5), sulfonamides (3), tetracycline (3), neuroleptic agents (1), triazene trypanocidal agents (1), other. (1)) in feeds. The proposed method with a modified Quick Polar Pesticides (QuPPe) sample preparation was validated for the determination of 30 veterinary drugs in feed samples by liquid chromatography triple-quadrupole mass spectrometry (LC-MS/MS). The sample was extracted with methanol containing 1% acetic acid and purified by dispersive solid-phase extraction (d-SPE) with C18. Good linearity (r2 ≥ 0.98) was observed, and the LOQ values ranged from 10 to 200 µg/kg. Average recoveries ranged from 70.8 to 118.4%, and the relative standard deviation was ≤ 18.7%. This validated method was used in the determination of 30 veterinary drugs in 142 feed samples obtained from South Korea. The results show that lincomycin was present in only one of the tested feed samples, although it was detected at a value lower than the LOQ. In conclusion, this multi-residue method can be used for screening through the detection and quantitation of residual multiclass veterinary drugs in feed samples.

In-House Validation of an Efficient and Rapid Procedure for the Simultaneous Determination and Monitoring of 23 Mycotoxins in Grains in Korea.

A high-performance liquid chromatography tandem mass spectrometry method is described for the simultaneous determination of mycotoxins, including Ergot alkaloids (EAs), in 3 types of grains. The extraction of 23 mycotoxins was evaluated and performed by using a modified QuEChERS-based sample preparation procedure. The proposed method was fully validated on spiked grain samples (barley, wheat and oat) to assess the linearity, limit of detection (LOD) and limit of quantitation (LOQ), matrix effects, precision and recovery. After validation, this method was applied to 143 samples of various types of 3 grains from the Republic of Korea to survey the level of mycotoxin contamination in Republic of Korean grains. A total of 42 grain samples (29%) were contaminated with at least one of these mycotoxins at levels higher than the LOQ. The results demonstrated that the procedure was suitable for simultaneously determining these mycotoxins in cereals and could be performed for their routine analysis in mycotoxin laboratories.

Multi-residue analytical method for detecting pesticides, veterinary drugs, and mycotoxins in feed using liquid- and gas chromatography coupled with mass spectrometry.

Modified QuEChERS and triple quadrupole mass spectrometry (LC and GC-MS/MS) technology were used to sequentially analyze pesticides, veterinary drugs, and mycotoxins in feed. In order to analyze the harmful substances that may remain or occur in the feed, we performed optimization experiments for sample preparation and LC-MS/MS and GC-MS/MS conditions. Optimized sample preparation involves extracting 5 g of sample with 15 mL of 0.25 M EDTA and 10 mL of acetonitrile. And some extracts were diluted 10-fold with 100 mM ammonium formate aqueous solution and analyzed by LC-MS/MS, and some extracts were purified through 25 mg PSA and analyzed by GC-MS/MS by adding an analyte protectant. We confirmed the matrix effect of feed ingredients and compound feeds, and added a dilution process after extraction to increase on-site efficiency. Matrix-matched calibration was applied for quantification. Method validation was performed for 197 pesticides, 56 components for veterinary drugs, and 5 components for toxins. All the components showed good linearity (r2 ≥ 0.98) in the developed analytical method. For most compounds, the limit of quantitation was 0.05 mg/kg. The recovery rate experiment was repeated three times at three concentrations including LOQ in feed ingredient, compound feed for livestock, and compound feed for pets. The recovery rate was 70.09-119.76% and relative standard deviations were ≤ 18.91%. And the accuracy and precision were further verified through cross-validation between laboratories. The developed analytical method was used to monitor 414 domestically distributed and imported feeds.

Structurally distorted perovskite La0.8Sr0.2Mn0.5Co0.5O3-δ by graphene nanoplatelet and their composite for supercapacitors with enhanced stability.

Supercapacitors are promising energy storage devices with high charging/discharging speeds and power densities. To improve their poor stability, we fabricated electrodes by integrating perovskite materials (La0.8Sr0.2Mn0.5Co0.5O3-δ, LSMCO) possessing redox reaction ability with graphene nanoplatelets exhibiting good electronic properties. One of the resultant composites (L25G70) demonstrated high capacitance and excellent capacitance retention (95% after 5000 cycles). These results are superior to other electrodes (L50G45 and L75G20) containing a larger ratio of LSMCO, even L75G20 did not exhibit supercapacitor behavior after 3000 cycles. GN can induce structural distortion in LSMCO, thereby the high amount of adsorbed oxygen per lattice oxygen can explain the best electrochemical performance of L25G70, while structural collapse rationalized the failure of L75G20. The findings of this study demonstrated that the use of LSMCO can improve the cycling stability of supercapacitors.

Robust Touch Screen Readout System to Display Noise Using Multireference Differential Sensing Scheme for Flexible AMOLED Display.

This paper presents a front-end architecture for touch screen panel (TSP) readout in a TSP-integrated, ultrathin flexible display to mitigate severe display noise interference, which is an uncommon mode caused by the large panel load of the TSP in the flexible display. The differential sensing method with multireference TSP channels minimized an imbalance of the phase and amplitude of the coupled-display noise interference. In addition, cascaded time-discrete bandpass sampling was employed to enhance the touch sensitivity in the sensing block. Moreover, a rated front-end block could be reconfigured to a differential or single-ended sensing structure, which reused the prefilter capacitors in the differential sensing for offset cancellation in reference capacitance sensing. To further improve the sensitivity, programmable postfiltering was employed on the reference TSP channels. Subsequently, the proposed front-end was implemented in a 350 nm process, wherein it achieved a SNR of 50.5 dB with a scan rate of 200 Hz and attenuated aggravated display noise interference by more than 6.84 dB as compared to the conventional differential sensing method. The designed chip occupied an area of 4.8 mm2 and consumed 17.6 mW from a 3 V supply.

Review of 107 Oncoplastic Surgeries Using an Acellular Dermal Matrix with the Round Block Technique.

The round block technique (RBT) is an oncoplastic surgery method that uses volume displacement techniques after partial mastectomy. However, cosmetic problems occur after tissue rearrangement in patients with small breasts or those in whom a large amount of breast tissue is excised. Therefore, we used an acellular dermal matrix (ADM) when the volume was insufficient after tissue rearrangement. Patients who underwent breast reconstruction using the ADM with the RBT after breast-conserving surgery (BCS) were included. The ADM graft was performed in two layers. First, it was placed on the glandular flap, and the patient was then seated to ascertain the degree of deformity. If the volume was insufficient, a graft was also performed under the skin flap. Overall, 107 oncoplastic surgeries were performed. Tumors were most commonly located in the upper outer quadrant of the breast, and the mean resected breast tissue was 27.1 g. Seroma was the most common complication, but it improved with several aspirations. There were no major complications or cosmetic problems requiring reoperation. Therefore, if the ADM was used for defects that could not be reconstructed with the RBT alone, safe and cosmetically good results could be obtained.

A monolithic linear motion platform driven by a piezoelectric and fluidic pressure-fed dual mechanism.

This paper presents a novel dual-mode motion mechanism capable of achieving nanopositioning on a monolithic linear motion platform. Unlike conventional dual-mode stages that use piezoelectric (PZT)- and electromagnetic-combined or similar actuation mechanisms comprising two separate motion axes, the dual-mode actuation was developed by combining a PZT for a coarse motion and a fluidic pressure-fed mechanism (FPFM) for a fine motion and was implemented in a monolithic flexure stage fabricated by metal additive manufacturing. The FPFM actuates the flexure stage by pressuring the media in the fluidic channels created inside the flexure spring structures. Experimental tests were performed to investigate the performance of the dual-mode linear motion platform. The stiffness, damping, and frequency response functions of the dual-mode stage were experimentally characterized. The proportional-integral-differential control combined with dual-mode control was employed to control the position of the flexure stage while bidirectionally controlling the flow of compressed air for a fine motion. The FPFM motion showed a good response to 1 nm stepwise input (every 10 psi), and it was implemented to provide up to ∼10 nm fine motion along with the PZT coarse motion (1 µm). The hysteresis characteristics of the FPFM were also characterized and compensated to track the positioning error.

Anti-diabetic effect of the lupinalbin A compound isolated from Apios americana: In vitro analysis and molecular docking study.

Dipeptidyl peptidase 4 (DPP4) and α-glucosidase inhibitors have been developed as anti-diabetic agents for the treatment of diabetes mellitus. In the present study, the anti-diabetic effects of the lupinalbin A compound isolated from Apios americana was investigated by measuring its inhibitory activity against DPP4 and α-glucosidase. To detect the inhibitory effect of lupinalbin A, DPP4 and α-glucosidase assays were performed in vitro. Molecular docking analysis was performed using AutoDock 4.2. The IC50 values of lupinalbin A against DPP4 and α-glucosidase were 45.2 and 53.4 µM, respectively. Analysis of the enzyme kinetics revealed that lupinalbin A interacted with the active site of DPP4 in a competitive manner, with an inhibition constant (Ki) value of 35.1±2.0 µM, whereas the lupinalbin A interaction with α-glucosidase was non-competitive, with a Ki value of 45.0 µM. Molecular docking analysis revealed a binding pose between the DPP4 enzyme and lupinalbin A. Taken together, these data suggest lupinalbin A is more effective against DPP4 than α-glucosidase, with regard to its anti-diabetic effects.

Anti-Arthritic Activities of Supercritical Carbon Dioxide Extract Derived from Radiation Mutant Perilla Frutescens Var. Crispa in Collagen Antibody-Induced Arthritis.

We investigated the anti-arthritic effects of the radiation mutant Perilla frutescens var. crispa leaf extract (SFE-M) and wild type leaf extract (SFE-W), both prepared by supercritical carbon dioxide (SC-CO2) extraction, on collagen antibody-induced arthritis (CAIA) in Balb/c mice. Animals were randomly divided into four groups: control, CAIA, CAIA + SFE-M (100 mg/kg/day), and CAIA + SFE-W (100 mg/kg/day). The mice were subjected to the respective treatments via oral gavage once daily for 4 days. Mice treated with SFE-M developed less severe arthritis than the CAIA mice. They showed significantly improved arthritic score, paw volume, and paw thickness compared to the CAIA mice from days 3 through 7. Furthermore, histopathological analysis of ankle for inflammation showed that SFE-M treatment reduced inflammatory cell infiltration and edema formation. Similarly, the neutrophil-to-lymphocyte ratio (NLR) in the whole blood was 37% lower in mice treated with SFE-M compared with the CAIA mice. However, treatment with SFE-W did not result in any significant difference compared with the CAIA group. In conclusion, SFE-M treatment delays the onset of arthritis and alleviates its clinical manifestations in CAIA mice.